Skip Navigation Links
 Venue 
 تاریخ های مهم 
 Registration 
 Pre Registered 
 Abstracts 
 برنامه همایش 
 Exhibition 
 Personal page 
  
 Archive 
                             نهمین همایش تحقیقات چشم پزشکی و علوم بینایی ایران       جشنواره فیلم       سومین همایش بهاره چشم پزشکی
Skip Navigation Links
        مکان برگزاری
        تاریخ های مهم
        ثبت نام
        ثبت نام شدگان
        مقالات
        برنامه همایش
        نمایشگاه
        صفحه شخصی
        جستجوی سخنران
        آرشیو
 
مقاله Abstract


Title: Generation of HEK-GIRK stable cell line as an applied model for optogenetic study
Author(s): Hoda Shams Najafabadi1, Zahra-Soheila Soheili1, Sharam Samiei3, Ehsan Ranaei Pirmardan4, Ali Kashanian5, Reza Salmanipour1, Hamid Ahmadieh6.
Presentation Type: Poster
Subject: Biochemistry/ Molecular Biology/Retinal Cell Biology
Others:
Presenting Author:
Name: Hoda Shams Najafabadi
Affiliation :(optional) Department of Molecular Medicine, National Institute of Genetic Engineering and Biotechnology, Tehran, Iran.
E mail: shams.hoda250@gmail.com
Phone:
Mobile: 0910201322
Purpose:

Optogenetic is a biological technique that uses light to restore neurons’ function in neurological disorders or to fix vision in visual impairment. Novel optogenetic tools belong to the GPCR protein (Opto-mGluR6) family that activates K+ channels in neural cells’ excitation. G protein-coupled inwardly-rectifying potassium channels (GIRKs) are a family of potassium ion channels which are activated via Opto-mGluR6. Therefore HEK293 cell line that stably express GIRK channels stands as a validated model for studying the principal optogenetic chimeric proteins. This study aims to generate stable GIRK expressing HEK293 cell line as a noteworthy model cell line.

Methods:

PcDNA3-GIRK1 and pcDNA3-GIRK2 were obtained as a gift from Dr. Terry Hebert, recovered transformed to XL10 E.coli. Plasmid mini preparation and subsequently sequencing experiments were performed, killing curve assay developed and optimal dose of G418 for HEK293 cell line was determined. HEK 293 cells were cotransfected with pcDNA3-GIRK1 and pcDNA3-GIRK2, maintained under G418 selection, and G418-resistant single cells were isolated. GIRKs expression in HEK-GIRK stable cell lines was assessed by RT-PCR.

Results:

GIRK1 and GIRK2 sequences were confirmed. 600 ug/ml was determined as optimal dose for G418 treatment. GIRK1 and GIRK2 plasmids efficiently co-transfected to HEK 293 cell line and resistant cells were detected. Through single cell isolation and expansion, expression of GIRK1 and GIRK2 were confirmed.

Conclusion:

HEK-GIRK stable cell line was generated as an applied model for future optogenetic study.

Attachment: 57Generation of HEK-GIRK stable cell line as an applied model for optogenetic study.pptx





Last News

  - نهمین همایش تحقیقات چشم پزشکی و علوم بینایی ایران
  - جشنواره فیلم
  - سومین همایش بهاره چشم پزشکی